KR20120032649A - Pharmaceutical composition for preventing or treating rheumatoid arthritis comprising vitis vinifera pip extract and rebamipide - Google Patents

Abstract

PURPOSE: A pharmaceutical composition containing rebamipide and Vitis vinifera seed extract is provided to prevent or treat rheumatoid arthritis. CONSTITUTION: A pharmaceutical composition for preventing or treating rheumatoid arthritis contains rebamipide and Vitis vinifera seed extract. The pharmaceutical composition is orally administered in the form of a tablet or capsule. The dosage of rebamipide is 0.3 mg/kg a day.

Description

Pharmaceutical composition for preventing or treating rheumatoid arthritis comprising Vitis vinifera pip extract and rebamipide}

The present invention relates to a pharmaceutical composition for the prevention or treatment of rheumatoid arthritis, including seed extract of European grapes and levamifeed as an active ingredient.

Rheumatoid arthritis is a chronic inflammatory disease characterized by inflammation and proliferation of synovial cells. Unlike osteoarthritis, osteoporosis and osteoporosis of bones around joints occur. Rheumatoid arthritis is a narrowing of the joints due to the gradual breakdown of the synovial membrane inflammation (joint capsule), ligaments, tendons (tendon), and joint cartilage. The tension of the joint membrane and the ligaments is lost (step 2), the inflammation invades the bones and causes partial erosion of the bones (step 3), and the joint function is lost (step 4).

Conventionally, for the treatment of rheumatoid arthritis, nonsteroidal anti-inflammatory agents such as NSAID, salicylate, COX-2 inhibitors; Corticosteroids such as prednisolone and triamcinolone; Anti-rheumatic drugs (DMARDs) such as methotrexate and sulfasalazine; And drugs such as biological agents such as etanercept, infliximab, and adalimumab. In general, non-steroidal anti-inflammatory agents and corticosteroids are used in the early stages of symptom expression, anti-rheumatic drugs are used in consideration of symptoms and disease activity, and in severe cases, biological agents or combination therapy are used in clinical practice.

However, biological agents have a problem in that they are difficult to apply in the medical field due to the high cost of treatment compared to existing therapeutic agents. Nonsteroidal anti-inflammatory drugs such as NSAID, which are relatively inexpensive to treat, are representative of gastrointestinal side effects, and antirheumatic drugs such as methotrexate and sulfasalazine are also pointed out as gastrointestinal disorders. Therefore, when the above drugs are applied to rheumatoid arthritis patients, cytoprotective agents such as levamipid, and H ₂ -receptor antagonists such as cimetidine and ranitidine are co-administered. Gastrointestinal disease occurs when an attacker (eg, stomach acid) is strong or when the defense factor is weak. Proton pump inhibitors such as omeprazole and gastric acid pump antagonists such as revaprazan are all compounds that inhibit the secretion of gastric acid, ie, sucralate and levamifeed, and mucoprotective agents such as sucralate and levamifeed are compounds that enhance the protective factor. The present inventors have completed the patent application by discovering that levamipid itself, which is used in combination to prevent gastrointestinal disorders, has a prophylactic and therapeutic activity against rheumatoid arthritis (Korean Patent Application No. 10-2009-0078547). , Undisclosed).

On the other hand, Vitis is a vine plant belonging to Rhamnales and Vitaceae, and is grown or grown all over the earth except near the equator and latitudes above 50 °. There are 700 genera of 11 genera, Vitis vinifera , Vitis labrusca , Vitis riparia , Vitis rupestris , Vitis rupestris , and Vitis berladieri , and Vitis. coignetiae ) and Vitis amurensis are mainly grown for fruit use. Among these, extracts obtained from seeds of the European species Vitis vinifera include (-) epicatechin, proanthocyanidins B1 and B2, (+) catechins, and mixtures of polymerized derivatives thereof, which are procyano Known as stone or flavonol oligomers (GB-A-1541469 and FR-A-2092743).

Seed extracts of the grape varieties Vitis vinifera mainly act on the glycosaminoglycans of connective tissues, blood vessels, lymph, joints, etc., to inhibit the synthesis of fibers related to the binding ability of collagen, elastin, fibronectin, etc. It has been reported to promote and inhibit degradation, inhibiting increased capillary permeability and restoring vein elasticity quickly. Arteres et veines Vol. 5 (5), 397-401 (1986); Sem-dex Hopitaux 48/47, 2009? 2013 (1981)] In addition, grape seed extract has been reported to have therapeutic effects on capillary vulnerability, hypertension-related retinopathy, and residual retinal edema after retinal detachment in diabetic patients. [Gazette Mde France Vol. 88, No. 14, 2035–2038 (1981)], which protects the various structures of the retina, speeds up the regeneration of visual pigment and is effective in restoring the retina after exposure to scintillation in clinical trials. Has been reported to be [Bull. Soc. Opth. France Vol 88 (2), 173-4, 177-9 (1988).

The present inventors and bars have developed an improved method for producing an extract of Vitis vinifera (Vitis vinifera), also, to see that the seed extract of the common grape vine (Vitis vinifera) has an excellent preventive and therapeutic effect on rheumatoid arthritis (Korean Patent Publication No. 10-2009-0024647).

The present inventors have found that the process of carrying out the prior studies of the inventors, that various tests on the basis of the lever is non-feed itself has a preventive and therapeutic activity against rheumatoid arthritis is prior research results, surprisingly, in the European species of grapes (Vitis vinifera) When a combination of seed extract and levamifeed is administered, the improvement or therapeutic activity of rheumatoid arthritis is further increased, that is, the synergistic effect prevents and treats rheumatoid arthritis for a long time (for example, 10 weeks or more). It was found that the activity can be maintained.

Accordingly, an object of the present invention is to provide a pharmaceutical composition for the prevention or treatment of rheumatoid arthritis, comprising as an active ingredient a seed extract of levamipid and European species Vitis vinifera .

According to one aspect of the present invention, there is provided a pharmaceutical composition for the prevention or treatment of rheumatoid arthritis, comprising a seed extract of levamipid and European species Vitis vinifera as an active ingredient.

In the pharmaceutical composition of the present invention, the pharmaceutical composition is preferably a pharmaceutical composition for oral administration, and the unit dosage form of the pharmaceutical composition is preferably a solid dosage form for oral administration in the form of a tablet or capsule.

According to the present invention, it was found that, when the seed extract of the grape varieties Vitis vinifera and levamifeed were administered in combination, the improvement or therapeutic activity of rheumatoid arthritis was further increased to show a synergistic effect. In other words, it has been found that the combination of levamipid and seed extracts of Vitis vinifera can maintain the prophylactic and therapeutic activity of rheumatoid arthritis for a long time (for example, 10 weeks or more). Therefore, the pharmaceutical composition containing seed extract and levamipid of European species grape ( Vitis vinifera ) as an active ingredient can be usefully used for the prevention or treatment of rheumatoid arthritis.

1 shows collagen-induced arthritis animals (CIA animals) and CIA animals with levamifeed ()) 0.3 mg / kg, seed extract of Vitis vinifera (Δ) 100 mg / kg, or European grapes. The results of arthritis evaluation were measured by oral administration of 100 mg / kg of seed extract of Vitis vinifera and 0.3 mg / kg of levamifeed.
FIG. 2 shows collagen-induced arthritis (CIA) animals and CIA animals with levamifeed (Reba 0.3 mg / kg), seed extract of European grape ( Vitis vinifera ) (GSPE 100 mg / kg), or European grape ( Vitis). Tissue staining of toxin and cartilage tissue destruction by killing animals orally administered with vinifera seed extract and levamipid (GSPE 100 + Reba 0.3). [Toluidine blue, safranin O] , H & E, and TRAP staining].

As used herein, the term "rebamipide" includes all forms of rebamipide, such as anhydrides, hydrates (e.g., half hydrates), crystalline forms, and the like. Acceptable salts. Pharmaceutically acceptable salts of the levamifeed are inorganic acid salts, acetic acid, formic acid, prepared from inorganic ion salts, hydrochloric acid, nitric acid, phosphoric acid, bromic acid, iodic acid, sulfuric acid, and the like prepared from calcium, potassium, sodium, magnesium, and the like. , Organic acid salts prepared from succinic acid, tartaric acid, citric acid, trichloroacetic acid, trifluoroacetic acid, gluconic acid, benzoic acid, lactic acid, fumaric acid, maleic acid, etc. Sulfonic acid salts prepared with toluenesulfonic acid and nathalenesulfonic acid and the like, amino acid salts made from glycine, arginine, lysine and the like, and amine salts made from trimethylamine, triethylamine, ammonia, pyridine, picoline, and the like. .

In addition, in the present invention, "seed extract of the grape varieties ( Vitis vinifera )" is a conventional manufacturing method using the seeds of the grape varieties ( Vitis vinifera ), for example, GB-A-1541469 and FR-A-2092743 What was obtained by the manufacturing method disclosed in can be used. Preferably, an improved production method developed by the inventors, for example, extracts obtained according to the production method disclosed in Korean Patent Publication No. 10-2009-0024647 can be used. The seed extract of the European species grape ( Vitis vinifera ) obtained by the improved manufacturing method is 80? Procyanidolic value (PCV) of 130; Content of (+) catechin and (−) epicatechin of up to 30%; And 95? It has a proanthocididine content of 105%.

The present invention provides a pharmaceutical composition for the prevention or treatment of rheumatoid arthritis, comprising the seed extract of levamipid and European grape ( Vitis vinifera ) as an active ingredient.

As can be seen in the following examples, the combination of levamipid and seed extract of Vitis vinifera in collagen-induced arthritis disease animals showed excellent rheumatoid arthritis therapeutic activity. In particular, the arthritis index was continuously suppressed when co-administered with the extract of levamifeed or Vitis vinifera , and the inhibitory activity was maintained during the 10-week trial period. (FIG. 1). The results of histological examination also showed a significantly lower degree of destruction of joints and cartilage compared to the group not administered or the group administered with the seed extract of levamifeed and Vitis vinifera in combination (FIG. 2). The results show that the pharmaceutical composition containing the seed extract and the levamipid of the European grape varieties ( Vitis vinifera ) as an active ingredient exhibits a synergistic effect, thereby having an excellent therapeutic activity for rheumatoid arthritis.

The pharmaceutical composition of the present invention may be administered orally or parenterally, preferably orally.

In the pharmaceutical composition of the present invention, the seed extract of levamifeed and Vitis vinifera is formulated in one unit dosage form (eg, in unit dosage form such as tablets, capsules, etc.) or separately It can be formulated in the unit dosage form of. The unit dosage form is preferably a solid dosage form for oral administration in the form of tablets or capsules.

The pharmaceutical composition of the present invention comprises a pharmaceutically acceptable carrier and is used for oral administration in the form of powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols, etc., preferably in the form of tablets or capsules according to conventional methods. It may be formulated into a solid dosage form. The pharmaceutically acceptable carrier is lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, microcrystalline cellulose , Hydroxypropyl cellulose, low-substituted hydroxypropyl cellulose, hydroxypropyl methyl cellulose 2910, polyethylene glycol 6000, polyvinyl pyrrolidone, methylhydroxy benzoate, propyl hydroxy benzoate, titanium oxide, talc, magnesium stearate Rate and mineral oil and the like. Also included are diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrants, surfactants, and the like. Dosage forms for oral administration of tablets, pills, powders, granules, capsules and the like include at least one excipient such as starch, calcium carbonate, sucrose or lactose, gelatin and the like. And may include lubricants such as magnesium stearate, talc, and the like. For example, a unit dosage form comprising levamifeed is, in addition to levamifeed, a low-substituted hydroxypropyl cellulose, microcrystalline cellulose, titanium oxide, hydroxypropylmethylcellulose 2910, polyethylene glycol 6000, hydroxy as a carrier. Tablet form, including propylcellulose and magnesium stearate. In addition, when formulated in a separate unit dosage form, the unit dosage form comprising levamipid may be in the form of a commercially available levamifeed-containing tablet (eg, costa tablets (Otsuka Pharmaceutical)) ; The unit dosage form comprising the seed extract of Vitis vinifera may be a solid dosage form for oral administration as disclosed in the prior art of the inventors, for example, Korean Patent Publication No. 10-2009-0024647.

The dosage of the seed extract of the levamifeed and Vitis vinifera contained in the pharmaceutical composition of the present invention varies depending on the condition and weight of the patient, the extent of the disease, the form of the drug, the route of administration, and the duration of time. Can be appropriately selected. For example, the levamipid can be administered orally at a dose of about 0.3 mg / kg per day, and the seed extract of the European grape ( Vitis vinifera ) can be administered orally at a dose of about 100 mg / kg per day. have. Of course, the dose is based on a preclinical study, and the dose that can be used clinically can be adjusted according to the condition and weight of the patient, the extent of the disease, the form of the drug, the route and duration of administration, and the like. As an example, as a formulation suitable for daily administration, the pharmaceutical composition of the present invention may comprise 150 mg to 300 mg of levamipids and 300 mg to 600 mg of seed extract of Vitis vinifera per unit formulation. Can be.

Hereinafter, the present invention will be described in more detail with reference to examples. However, these examples are for illustrating the present invention, and the scope of the present invention is not limited to these examples.

Example. Evaluation of Rheumatoid Arthritis Treatment Efficacy Following Oral Administration

Mice inducing rheumatoid arthritis by administering collagen type 2 were prepared, and after oral administration with levamifeed , seed extract of Vitis vinifera , and these, the arthritis index was evaluated for 10 weeks. In addition, after 10 weeks, each mouse was killed and the degree of destruction of joint and cartilage tissue was observed by tissue staining (toluidine blue, safranin O, H & E, and TRAP staining). In the following examples, the seed extract of the European species grape ( Vitis vinifera ) used the extract obtained according to the prior art of the inventors, namely Example 1 of Korean Patent Publication No. 10-2009-0024647.

1. Preparation and Administration of Animal Models

The test animals used male DBA / 1J mice 6-6 weeks old. Type 2 collagen (CII) is dissolved in 0.1N acetic acid solution to 4 mg / ml, and then dialyzed with dialysis buffer (dialysis buffer, 50mM Tris, 0.2N NaCl) to complete M. tuberculosis. The mixture was mixed in the same amount with Freud's adjuvant (CFA, Chondrex) and injected subcutaneously into the base of the tail of the mouse to inject 50 μl per animal (ie 100 μg / 50 μl) (primary injection). Two weeks later, 4 mg / ml of bovain type II collagen was mixed 1: 3 with sterile PBS and injected with 100 μl (ie, 100 μg / 100 μl) of DBA / 1J (ie, 100 μg / 100 μl) (second injection). After the second injection, 0.3 mg / kg dose of levamipid, 100 mg / kg dose of Vitis vinifera seed extract, or a mixture thereof was orally administered using an oral zone. Oral administration was performed 10 times, once every two days for 20 days. Six groups of animals were used in each group. Arthritis was evaluated up to 10 weeks. After 10 weeks, each animal was killed to investigate the activity of rheumatoid arthritis in the blood and joint tissues.

2. Evaluation of Rheumatoid Arthritis Therapeutic Activity in CIA Animals

(2-1) Average joint index evaluation

Two weeks after the first inoculation, three observers, who did not know the contents of the experiment, evaluated the severity of joint inflammation three times a week and observed it for up to 10 weeks. At this time, the evaluation of arthritis is based on the average arthritis index by Rossoliniec et al., The average score divided by 3 is obtained by adding the scores according to the following scales on the three legs except the legs to which CII / CFA was administered at the time of the second dose per horse. The mean obtained by summing the values obtained by three observers in the animal model was used. The scores and criteria according to arthritis evaluation are as follows.

0 points: No lack or swelling.

1 point: Mild swelling and redness limited to the foot or ankle joint

2 points: Mild swelling and redness from the ankle joint to the metatarsal

3 points: moderate swelling and redness from the ankle joint to the ankle bone

4 points: swelling and redness from the ankle to the entire leg

The best arthritis index per head is 4, so the best disease index per rat is 16. The arthritis index was suppressed initially (approximately 4 weeks) in the group orally administered levamipid at a dose of 0.3 mg / kg and the group orally administered the seed extract of Vitis vinifera at 100 mg / kg. This showed a therapeutic activity of rheumatoid arthritis, but significantly increased arthritis index after about 5 weeks. This indicates that the prophylactic or therapeutic activity of rheumatoid arthritis at this dose is not satisfactory. On the other hand, the combination of levamifeed and seed extracts of Vitis vinifera co-administered the arthritis index continuously, and this inhibitory activity was maintained for 10 weeks of testing (FIG. 1). .

(2-2) Histological Examination

After 10 weeks of killing the test animals, the hind paws of the mice were fixed with 10% formalin, decalcified from bone, and paraffin-embedded. Joint sections (5-7um) were prepared and stained with hematoxylin and eosin. In addition, histological examination was performed with toluidine blue, safranin O, and TRAP staining to confirm the degree of cartilage destruction.

As a result of histological examination, the joints of CIA animals were infiltrated with many immune cells, and the formation of pannus, cartilage destruction and bone erosion were observed. On the other hand, animals treated with the seed extracts of levamifeed and Vitis vinifera in combination maintained the joint and cartilage destruction similar to those of normal mice, compared with normal mice. Much similar (FIG. 2).

Therefore, the test results indicate that when combined administration of the seed extract of levamipid and grape varieties ( Vitis vinifera ), it has an excellent preventive and therapeutic activity against rheumatoid arthritis by inhibiting the destruction of cartilage.

Claims (3)

A pharmaceutical composition for the prevention or treatment of rheumatoid arthritis, comprising a seed extract of levamipid and European species Vitis vinifera as an active ingredient. The pharmaceutical composition of claim 1, wherein the pharmaceutical composition is a pharmaceutical composition for oral administration. The pharmaceutical composition according to claim 2, wherein the unit dosage form of the pharmaceutical composition is a solid dosage form for oral administration in the form of a tablet or capsule.

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